Abstract：
BACKGROUND: Long non-coding RNA (lncRNA) LINC00460 is an onco-lncRNA in a variety of cancers, including pancreatic cancer (PC). This study is aimed to investigate the regulatory mechanisms of LINC00460 in PC.
METHODS: The tumor and adjacent normal tissues were collected from 73 PC patients. The expression of LINC00460, miR-503-5p, and ANLN was detected using qRT-PCR. We then analyzed the proliferation, migration, invasion, and apoptosis/cell cycle of PC cells by performing the MTT/EdU, transwell, and flow cytometry assays, respectively. The xenograft tumor model were utilized to confirm the effect of LINC00460 knockdown on PC through anti-PD-1 therapy in vivo, and the sensitivity of PANC-1 cells to the cytotoxicity of CD8+ T cells in vitro. Western blotting was used to determine the protein levels. A co-culture model was utilized to explore the effects of exosomes on macrophages.
RESULTS: LINC00460 was up-regulated in PC tissues and cells. LINC00460 knockdown suppressed cell proliferation, migration, and invasion, facilitated cell apoptosis and G0/G1 phase arrest, and inhibited the tumor growth through anti-PD-1 therapy. Both miR-503-5p down-regulation and ANLN up-regulation reversed the effects of LINC00460 knockdown on inhibiting the proliferation, migration and invasion, and on promoting the apoptosis, G0/G1 phase arrest, and the sensitivity of PC cells to the cytotoxicity of CD8+ T cells. Exosomes were uptaken by the ambient PC cells. PANC-1 cells-derived exosomal LINC00460-induced M2 macrophage polarization accelerates the cell migration and invasion.
CONCLUSIONS: LINC00460 silencing attenuates the development of PC by regulating the miR-503-5p/ANLN axis and exosomal LINC00460-induced M2 macrophage polarization accelerates the migration and invasion of PANC-1 cells, thus LINC00460 may act as a possible therapeutic target for treating PC.
METHODS: The tumor and adjacent normal tissues were collected from 73 PC patients. The expression of LINC00460, miR-503-5p, and ANLN was detected using qRT-PCR. We then analyzed the proliferation, migration, invasion, and apoptosis/cell cycle of PC cells by performing the MTT/EdU, transwell, and flow cytometry assays, respectively. The xenograft tumor model were utilized to confirm the effect of LINC00460 knockdown on PC through anti-PD-1 therapy in vivo, and the sensitivity of PANC-1 cells to the cytotoxicity of CD8+ T cells in vitro. Western blotting was used to determine the protein levels. A co-culture model was utilized to explore the effects of exosomes on macrophages.
RESULTS: LINC00460 was up-regulated in PC tissues and cells. LINC00460 knockdown suppressed cell proliferation, migration, and invasion, facilitated cell apoptosis and G0/G1 phase arrest, and inhibited the tumor growth through anti-PD-1 therapy. Both miR-503-5p down-regulation and ANLN up-regulation reversed the effects of LINC00460 knockdown on inhibiting the proliferation, migration and invasion, and on promoting the apoptosis, G0/G1 phase arrest, and the sensitivity of PC cells to the cytotoxicity of CD8+ T cells. Exosomes were uptaken by the ambient PC cells. PANC-1 cells-derived exosomal LINC00460-induced M2 macrophage polarization accelerates the cell migration and invasion.
CONCLUSIONS: LINC00460 silencing attenuates the development of PC by regulating the miR-503-5p/ANLN axis and exosomal LINC00460-induced M2 macrophage polarization accelerates the migration and invasion of PANC-1 cells, thus LINC00460 may act as a possible therapeutic target for treating PC.
摘要：
背景：长非编码RNA（lncRNA）LINC00460是多种癌症中的onco-lncRNA，包括胰腺癌（PC）。本研究旨在探讨LINC00460在PC中的调控机制。
方法：收集73例PC患者的肿瘤及癌旁正常组织。LINC00460,miR-503-5p,使用qRT-PCR检测ANLN。然后我们分析了增殖，迁移,入侵,通过MTT/EdU进行PC细胞的凋亡/细胞周期，transwell,和流式细胞术检测，分别。异种移植肿瘤模型用于确认LINC00460敲低通过体内抗PD-1治疗对PC的影响。以及PANC-1细胞对体外CD8+T细胞毒性的敏感性。使用蛋白质印迹法测定蛋白质水平。利用共培养模型来探索外泌体对巨噬细胞的影响。
结果：LINC00460在PC组织和细胞中上调。LINC00460敲低抑制细胞增殖，迁移,和入侵,促进细胞凋亡和G0/G1期阻滞，并通过抗PD-1治疗抑制肿瘤生长。miR-503-5p下调和ANLN上调均逆转了LINC00460敲低对细胞增殖的抑制作用,移民和入侵，促进细胞凋亡，G0/G1相阻滞,和PC细胞对CD8+T细胞的细胞毒性的敏感性。外泌体被周围的PC细胞摄取。PANC-1细胞来源的外泌体LINC00460诱导的M2巨噬细胞极化加速细胞迁移和侵袭。
结论：LINC00460沉默通过调节miR-503-5p/ANLN轴减弱PC的发育，外泌体LINC00460诱导的M2巨噬细胞极化加速PANC-1细胞的迁移和侵袭，因此LINC00460可以作为治疗PC的可能的治疗靶标。
方法：收集73例PC患者的肿瘤及癌旁正常组织。LINC00460,miR-503-5p,使用qRT-PCR检测ANLN。然后我们分析了增殖，迁移,入侵,通过MTT/EdU进行PC细胞的凋亡/细胞周期，transwell,和流式细胞术检测，分别。异种移植肿瘤模型用于确认LINC00460敲低通过体内抗PD-1治疗对PC的影响。以及PANC-1细胞对体外CD8+T细胞毒性的敏感性。使用蛋白质印迹法测定蛋白质水平。利用共培养模型来探索外泌体对巨噬细胞的影响。
结果：LINC00460在PC组织和细胞中上调。LINC00460敲低抑制细胞增殖，迁移,和入侵,促进细胞凋亡和G0/G1期阻滞，并通过抗PD-1治疗抑制肿瘤生长。miR-503-5p下调和ANLN上调均逆转了LINC00460敲低对细胞增殖的抑制作用,移民和入侵，促进细胞凋亡，G0/G1相阻滞,和PC细胞对CD8+T细胞的细胞毒性的敏感性。外泌体被周围的PC细胞摄取。PANC-1细胞来源的外泌体LINC00460诱导的M2巨噬细胞极化加速细胞迁移和侵袭。
结论：LINC00460沉默通过调节miR-503-5p/ANLN轴减弱PC的发育，外泌体LINC00460诱导的M2巨噬细胞极化加速PANC-1细胞的迁移和侵袭，因此LINC00460可以作为治疗PC的可能的治疗靶标。
